Development of chromosome sorting techniques using a dual laser chromosome sorter

使用双激光染色体分选机开发染色体分选技术

• 批准号: 61880023
• 负责人: MATSUBARA Kenichi
• 金额: $ 23.49万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research
• 财政年份: 1986
• 资助国家: 日本
• 起止时间: 1986 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-61880023/
• 关键词: Cell Sorter; Dual Laser; 染色体

We have developed chromosome sorting techniques using a dual laser chromosome sorter to analyze the structure of human chromosomes.Human chromosomes stained with Hoechst 33258 (Ho) and chromomycin A3 (CA3) flowed sequentially through two laser beams, one adjusted to 457.9nm to excite CA3 and the other adjusted to 351.1-363.8nm to excite Ho. The Ho and CA3 contents of individual chromosomes were determined by measuring the intensities of fluorescence that were emitted as they passed two laser beams. These measurements were accumulated to form fluorescence intensity distributions.Sorting techniques have evolved from the ability to sort 10-20 human chromosome fractions with a single laser system to sorting 21 fractions including 15 unique types with a dual laser system.dFurthermore, chromosome types that cannot be sorted at high purity from chromosomes isolated from human cells can be sorted from human-hamster hybrid cell lines containing one or a few human chromosomes.

我们用双激光染色体分选仪分析了人类染色体的结构，用Hoechst 33258（Ho）和色霉素A_3（CA_3）染色的人类染色体依次通过两束激光，一束激光激发CA_3，另一束激光激发Ho，波长分别为457.9nm和351.1-363.8nm。单个染色体的Ho和CA 3含量通过测量当它们通过两个激光束时发射的荧光强度来确定。分选技术已经从用单个激光系统分选10-20个人类染色体片段的能力发展到用双激光系统分选包括15种独特类型的21个片段。不能从分离自人细胞的染色体中以高纯度分选的染色体类型可以从人-含有一条或几条人类染色体的仓鼠杂交细胞系。

项目成果

Horii,Akira: "Primary structure of human pancreatic secretory trypsin inhibitor (PSTI) gene" Biochem.Biophys.Res.Commun.149. 635-641 (1987)

堀井晃：“人胰腺分泌型胰蛋白酶抑制剂（PSTI）基因的一级结构”Biochem.Biophys.Res.Commun.149。

Tokino,Takashi: "Chromosomal translocation and inverted duplication associated with integrated hepatitis B virus in hepatocellular carcinomas" J.Virol.61. 3848-3854 (1987)

Tokino，Takashi：“肝细胞癌中与整合型乙型肝炎病毒相关的染色体易位和反向重复”J.Virol.61。

福重真一: 続生化学実験講座1 遺伝子研究法【III】(日本生化学会編). 301-313 (1987)

Shinichi Fukushige：后续生物化学实验课程1基因研究方法[III]（日本生化学会编辑）301-313（1987）。

Horii,Akira: Biochem.Biophys.Res.Commun.149. 635-641 (1987)

堀井晃：Biochem.Biophys.Res.Commun.149。

Fukushige,Shinichi: "Chromosomal assignment of human genes for gastrin, thyrotropin (TSH)- subunit and c-erbB-2 by chromosome sorting combined with velocity sedimentation and Southern hybridization" Biochem.Biophys.Res.Commun.134. 477-483 (1986)

Fukushige,Shinichi：“通过染色体分选结合速度沉降和 Southern 杂交对胃泌素、促甲状腺素 (TSH) 亚基和 c-erbB-2 人类基因进行染色体分配”Biochem.Biophys.Res.Commun.134。